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Gelsolin TL40 (N-terminal cytoplasmic gelsolin) - 50µg

Cat. #:
8305-01
Shipping:
FedEx Express (Global) FedEx Express (Global) (shipping info)
Quantity:
50 µg




TL40 (N-terminal cytoplasmic gelsolin)
(Actin Binding Protein)


Cat. #: 8305-01  

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Applications

  • Fragmentation assays in the absence of Ca++ (permits the presence of EGTA);
  • Easy and efficient removal of F-actin from the cellular or myofibrillar cytoskeleton;


Description

   

Protein

TL40 (N-terminal cytoplasmic gelsolin)

Origin

smooth muscle, porcine

Molecular mass

40kDa

Protein description

TL40 was obtained by thermolysin cleavage of smooth muscle gelsolin. It consists of the N-terminal homologous subdomains S1-S3 (aa 1-361, molacular mass: ~40kD) and is a Ca++-independent proteolytic fragment of native gelsolin.

TL40 is preferentially used in F-actin fragmentation assays in the presence of EGTA.

Actin interaction

Ca++-insensitive F-actin severing.


Properties

   

Form

Lyophilized, ready-to-use.

Quantity

1x50 µg

Buffer

Lyophilized TL40 (50µg) contains 20mM imidazole pH 7.2, 50mM KCl, 1.0mM EGTA and 0.1mM DTT and 0.5% disaccharides,  when reconstituted with 50µl ultrapure water to obtain a 1.0 mg/ml solution.

Purity & Activity

 

Purity: >98% by scanning densitometry from Coomassie G-250 stained SDS-Gels.

Fragmentation activity: The fragmentation activity of TL40 was determined by Ostwald viscometry at 25°C. The specific viscosity of F-actin (1mg/ml) is reduced by <45% after addition of TL40 at a molar ratio of 1 : 100 (TL40 : A) within less than 2min.

For this purpose the TL40 stock solution was used as described below containing 1% disaccharides. F-actin (100mM KCl, 10mM Imidazole pH 7.4, 1mM ATP) was mixed with TL40 and instantly measured within a time course of 0-30min by Ostwald viscometry. 

Purification notes

GPC, IEXC.

Protein concentration

Determined by Biuret method with actin (skeletal muscle, rabbit) as reference.

Storage instructions

Gelsolin  is stored at –70°C upon arrival will be stable in performance for at least 6 months from the date of purchase. The solubilized protein is kept on ice and should be used within 5 days. Storage in glycerol at -20°C results in a significant decrase of the activity, and has to determined individually. 

Shipping conditions 

At ambient temperature. Upon delivery store at -70°C.

Remarks

For Use in Research only. Not for Use in Human or Veterinary Diagnostical or Therapeutical Applications. 

CAS no.

 





Further Information

Product DataSheet

Material and Safety Data Sheet


Protein Sequence on NCBI


Protein Structure on NCBI (PDB ID: 1D0N)

Please note that the structure PDB ID 1D0N is identical with cytoplasmic gelsolin, since it lacks the N-terminal extension of plasma gelsolin used for crystallization.



 

References

 

Ca2+ binding by domain 2 plays a critical role in the activation and stabilization of gelsolin.
Nag S, Ma Q, Wang H, Chumnarnsilpa S, Lee WL, Larsson M, Kannan B, Hernandez-Valladares M, Burtnick LD, Robinson RC.
Proc Natl Acad Sci U S A. 2009 Aug 18;106(33):13713-8. 

 

The crystal structure of plasma gelsolin: implications for actin severing, capping, and nucleation.
Burtnick LD, Koepf EK, Grimes J, Jones EY, Stuart DI, McLaughlin PJ, Robinson RC.; Cell. 1997 Aug 22;90(4):661-70.

 
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